We tried that trick, which works amazingly well in insect cells, in
mammalian media, and it fails. It will depend on the exact media, obviously.
Engin
On 10/8/09 1:50 PM, Matthew Franklin wrote:
> The trick we used at Genentech (which I'm still using) was for
> secreted insect cell proteins, but it should work for you as well. Add
> 1 mM NiCl2 and 10 mM CaCl2 to your conditioned media, and adjust the
> pH to 7.2 - 7.5. For insect cell conditioned media, this produces a
> fairly heavy precipitate which appears to contain the nickel-chelating
> factor. Most of the time (but not always!) your protein of interest
> will not be precipitated by this step, and you can spin out the
> precipitate then load the supernatant on to your nickel column with no
> further processing.
> Hope that helps,
>
> Matt
>
>
> --
> Matthew Franklin , Ph.D.
> Senior Scientist, ImClone Systems,
> a wholly owned subsidiary of Eli Lilly& Company
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> Thank you.
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--
Engin Özkan
Post-doctoral Scholar
Howard Hughes Medical Institute
Dept of Molecular and Cellular Physiology
279 Campus Drive, Beckman Center B173
Stanford School of Medicine
Stanford, CA 94305
ph: (650)-498-7111
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